Andrographolide induces cell cycle arrest and apoptosis in human rheumatoid arthritis fibroblast-like synoviocytes.

The pseudo-tumoral expansion of fibroblast-like synoviocytes is a hallmark of rheumatoid arthritis (RA), and targeting rheumatoid arthritis fibroblast-like synoviocytes (RAFLSs) may have therapeutic potentials in this disease. Andrographolide, a diterpenoid compound isolated from the herb Andrographis paniculata, has been reported to have potent anti-inflammatory activity. In the present study, we aimed to investigate the effects of andrographolide on human RAFLSs and the underlying molecular mechanism(s). RAFLSs were isolated from patients with RA and treated with or without various concentrations (i.e., 10, 20, and 30 µM) of andrographolide for 48 h. 3-[4,5-Dimethyl-2-yl]-2,5-diphenyl tetrazolium bromide assay revealed that andrographolide treatment decreased the proliferation of RAFLSs in a dose-dependent manner. Cell cycle analysis using propidium iodide (PI) staining showed a G0/G1 cell cycle arrest in andrographolide-treated RAFLSs. Immunoblotting analysis of key cell cycle regulators demonstrated that andrographolide treatment caused a dose-dependent increase in the expression of cell-cycle inhibitors p21 and p27 and a concomitant reduction of cyclin-dependent kinase 4. Exposure to andrographolide-induced apoptosis of RAFLSs measured by annexin V/PI double staining, which was coupled with promotion of cytochrome C release from mitochondria and activation of caspase-3. Moreover, andrographolide-treated RAFLSs displayed a significant decrease in the Bcl-2/Bax ratio compared to untreated cells. In conclusion, our data demonstrate that andrographolide exerts anti-growth and pro-apoptotic effects on RAFLSs, thus may have therapeutic potential for the treatment of RA.

[Expression of fas protein of myocardium in dilated cardiomyopathy].

OBJECTIVE: To investigate Fas protein expression of the myocardium in dilated cardiomyopathy (DCM) and its relationship with occurrence of sudden death caused by DCM. METHODS: Nine autopsy cases of sudden death caused by DCM along with the heart samples were chosen from the archives in the Department of Forensic Medicine, Tongji Medical College, HUST from 1997 to 2007. Other 11 cases which died of violence and other diseases were selected as the control group. Expressions of myocardial Fas protein in the samples were quantitatively detected by immunohistochemistry and computerized imaging analysis. RESULTS: Myocardial Fas protein expression increased significantly in the DCM group. Positive color showed brown-yellow granulated or striped distribution in the longitudinal section of myocardial within the cell membrane and cytoplasm, and showed circular brown granules in the cross section of the cell membrane, while these changes were not observed in the control group though there was focal weak staining noted. Statistical significance was observed between the experimental and control groups (P = 0.002), but no statistical significance was found for the average optical density value between these two groups (P = 0.675). CONCLUSION: The expression of Fas protein increased obviously in the DCM group. Such alteration in expression quantity and distribution of myocardial Fas protein may be related to arrhythmia and heart failure in the patients with DCM.

[Toll-like receptor 4 expression in the coronary atherosclerotic plaques of patients died from sudden cardiac death].

OBJECTIVE: To investigate the Toll like receptor 4 (TLR4) expression in the coronary atherosclerotic plaques in patients died from sudden cardiac death (SCD) or non SCD. METHODS: Autopsied coronary artery samples from 75 patients died from SCD (n = 28), non-SCD (n = 28) or non-CHD (n = 19) were examined and the R value (positive cells' areas/scanning areas) and A value (average optical density) of TLR4 expression in the coronary arteries were detected qualitatively by the immunohistochemistry (SABC method) and image analysis technologies. RESULTS: SCD group: 13 (46.4%) cases showed strong positive expression of TLR4; 11 (39.3%), positive expression; 4 (14.3%), weak positive expression. CHD group: 8 (28.6%) cases showed weak positive expression; 17 (60.7%), very weak positive expression; 3 (10.7%), no positive expression. There was no positive expression of TLR4 in non-CHD samples. A (1.140 +/- 0.101) and R value (0.0269 +/- 0.0027) in SCD group were significantly higher than in non-SCD and control groups (all P < 0.01). A value was siginificantly higher in CHD group (0.719 +/- 0.205) than that in control group (0.481 +/- 0.033, P < 0.05) while R value (0.0085 +/- 0.0007, 0.0046 +/- 0.0004) was similar between the groups (P > 0.05). CONCLUSION: The increased positive expressive of TLR4 in the atherosclerotic plaque can be regarded as an important pathological marker of SCD.

[Recent advances in venous air embolism].

Air embolism, a life-threatening complication of medical procedure, is a frequently encountered challenge in the forensic practices. It can be easily missed due to ignorance by forensic examiner or it could be difficult to be identified due to prolonged storage of the cadaver. This article reviews the etiological factors, pathophysiological changes, clinical manifestation, diagnosis, and the medicolegal identification of air embolism. The authors suggest that modern imaging techniques including echocardiogram, computer tomography, and magnetic resonance imaging play animportant role in the clinical diagnosis and forensic identification of air embolism.

[Pathomorphologic study of the fibrous cap and fatty core of coronary arthrosclerotic plaque in sudden coronary death].

OBJECTIVE: The differences in the thickness of fibrous cap and the percentage of fatty core of the coronary atherosclerotic plaques between sudden coronary death (SCD) group and the control group were investigated. METHODS: Sixty-four autopsy cases were divided into SCD and control groups. Samples were taken from the most severely damaged portions of the coronary atherosclerotic plaques, sectioned, stained with HE, and the percentage of examined by light microscopy for morphologic changes and structural alternations. Image analysis system was adopted to compare the thickness of fibrous cap and percentage of fatty core in the whole plaque between the two groups, and allthe data were analyzed and calculated with SPSS 11.5 statistic software. RESULTS: There were 15 grade III and 21 grade IV atherosclerotic cases found in the SCD group, while there were 16 and 12 found in the control group, respectively. Although no significant differences on the severity of atherosclerosis were found between the two groups (P > 0.05), there were significant differences on the thickness of the fibrous cap and the percentage of fatty core found between the two groups (P < 0.01). CONCLUSION: Our study indicates that there are significant differences in the thickness of fibrous cap and the percentage of fatty core in atherosclerosis plaques between the SCD group and the control group. These observed differences may be helpful for morphological diagnosis of SCD.

[Monocyte chemotactic protein-1 expression in coronary atherosclerosis plaque of sudden coronary death patients].

OBJECTIVE: To investigate the expression of monocyte chemotactic protein 1 (MCP-1) in coronary atherosclerosis plaque of sudden coronary death (SCD) patients and the relationship between MCP-1 expression and SCD. METHODS: Autopsy heart samples (n = 90) collected during 2001 - 2003 were divided to SCD group (n = 36) and 2 control groups: control group I, non-SCD CHD (n = 28), control group II, non-cardiac death (n = 26). The immuno-histochemistry SABC techniques (R, positive MCP-1 cell area/totl area) and computerized images analysis (A) were performed to detect the expression of MCP-1 in different groups. RESULTS: R and A in plaques are significant higher in SCD group than control group I and II (0.1264 +/- 0.013 vs 0.0269 +/- 0.0110 and 0.0267 +/- 0.0100, P = 0.04), (0.4534 +/- 0.083 vs 0.2303 +/- 0.040 and 0.2158 +/- 0.0400, P = 0.00), and similar between control group I and control group II. CONCLUSIONS: MCP-1 expression is increased in coronary atherosclerosis plaque of SCD patients.

Determination of free salicylic acid in chewing aspirin tablets by HPLC.

OBJECTIVE: To establish a HPLC method for determining the content of free salicylic acid in chewing aspirin tablets. METHOD: The determination was conducted on a HPLC column (C(18), 150 mm x 4.6 mm x 5 microm) with methanol-water-glacial acetic acid (8.0 5.5 1.0) as the mobile phase and the detection wavelength of 302 nm. RESULTS: The calibration curve was linear within the concentration range of 2.65 to 31.77 microg/ml (r=0.999 97) of salicylic acid. The average recovery rate was 100.21% with relative standard deviation of 0.53% (n=6). CONCLUSION: HPLC is quick and accurate of determining the content of free salicylic acid for chewing aspirin tablets.

[Progression of the manner of cardiomyocyte death and its potential significance in forensic medicine].

The manner of cell death is a hotspot of medical researchers. Apoptosis and necrosis were considered as two manners of cell death in the past. But recently a new manner of cell death--oncosis is gradually accepted by the pathologists. Oncosis is different from apoptosis in morphologic, mechanism and the role in cardiovascular diseases. In this paper, the progression of the research about manner of the cardiomyocyte death and its significance in forensic medicine in recent years was reviewed.

[The advance of protection for hazard factor during autopsy].

Recently, the special characteristics of work with SARS require particular attention to the facilities, equipment, policies and procedures involved. In fact, an autopsy also subject prosectors and others to a wide variety of hazards, including bloodborne, aerosolized pathogens and others (for example SARS). Forensic pathologists and other persons in close proximity to an autopsy need personal protective equipment, fourthemore, laboratory procedure and facility design principles of biosafety should be established for the protection of all personnal involved in the work.

[Gap junctions, connexins and sudden death caused by coronary heart disease].

Gap junctions construct hydrophilic trans-membrane channels which adjust the intercellular communication of chemistry and electricity. In the heart, individual cardiac myocytes are linked by gap junctions. These junctions form low resistance pathways along which the electrical impulse flows rapidly and repeatedly between all the myocardium, ensuring their synchronous contraction. In recent years, some researchers have found that connexins, the protein molecules of gap junction channels, are reduced in number or redistributed from intercalated disks (ID) to lateral cell borders in a variety of cardiac disease, especially in ischemic heart disease. The gap junction remodeling is considered to be arrhythmogenic. These findings will lead us to a new realm in the diagnostic of sudden death caused by coronary heart disease.

[Determination of isoniazid in blood and urine samples by reversed-phase high performance liquid chromatography].

For the purpose of more accurate and rapid analysis of isoniazid in body fluid samples, a reversed-phase high performance liquid chromatographic (HPLC) method was developed. Vanillin, being as a derivatizing reagent, was added to the plasma and urine samples when they were pretreated. Isoniazid and vanillin reacted to form isonicotinoyl hydrazone which was separated and detected. The pretreatment method of sample, the linear range, the precision, and the recovery of isoniazid were investigated by using human's plasma and urine spiked with standard isoniazid. The linear range was 0.2 mg/L- 12.0 mg/L (for plasma, r =0.999 6; for urine, r = 0.999 4). The detection limit was 0.2 mg/L. The intra-day and inter-day precisions of assay for isoniazid were 2.3% - 2.6% and 3.8% - 4.0% (n = 5) for plasma, 1.3% - 4.0% and 2.1% - 3.9% (n = 5) for urine respectively. The average recoveries of isoniazid were from 96.5% to 99.8% in plasma and from 96.3% to 99.4% in urine. The HPLC method has been used to investigate the concentration of isoniazid in the volunteer's plasma. The quantitative analysis of isoniazid in plasma was not interfered by impurities, the metabolites of isoniazid and the derivatizing reagent residue. This analytical method for isoniazid is sensitive, rapid and convenient. It is suitable for the analysis of toxics in forensic samples, the detection of drug concentration in clinical medicines and pharmacokinetic studies.